<p>U okviru ove disertacije ispitane su mogućnosti primene savremenih metoda za dobijanje tečnih ekstrakata (ultrazvučna ekstrakcija, mikrotalasna ekstrakcija i ekstrakcija subkritičnom vodom) i lipofilnih ekstrakata (ekstrakcija superkritičnim ugljendioksidom) bogatih polifenolima iz nadzemnog dela očajnice (Marrubium vulgare L.). Tečni ekstrakti su optimizovani primenom eksperimentalnog dizajna i metode odzivne povr&scaron;ine, i podvrgnuti su spray drying tehnici su&scaron;enja. Tečni i suvi ekstrakti su detaljno ispitani po pitanju hemijskog sastava i biolo&scaron;ke aktivnosti, antimikrobne i in vitro antihiperglikemijske aktivnosti.<br />Za svaku od navedenih ekstrakcionih tehnika ispitan je uticaj glavnih parametara: ultrazvučna ekstrakcija (temperatura, vreme ekstrakcije i snaga ultrazvuka), mikrotalasna ekstrakcija (koncentracija etanola, vreme ekstrakcije i snaga mikrotalasa) i ekstrakcija subkritičnom vodom (temperatura, vreme ekstrakcije i koncentracija HCl u ekstragensu). Kod tečnih ekstrakata dobijenih ultrazvučnom i mikrotalasnom ekstrakcijom na optimizovanim uslovima utvrđeno je prisustvo hlorogenske kiseline i kvercetina koji nisu detektovani u maceratima. Ispitana je antimikrobna aktivnost oba ekstrakta i utvrđeno da je B. cereus najosetljiviji bakterijski soj, dok je u slučaju kvasaca S. cerevisiae posebno osetljiv na oba ekstrakta. Pored ove aktivnosti ispitana je i in vitro antihiperglikemijska aktivnost oba ekstrakta prema &alpha;-amilazi i &alpha;-glukozidazi, pri čemu ultrazvučni ekstrakt pokazuje veću antihiperglikemijsku aktivnost (50,63% inhibira &alpha;-amilazu i 48,67% inhibira &alpha;-glukozidazu) &scaron;to se potencijalno može objasniti prisustvom hlorogenske kiseline (33,11 &mu;g/mL) i kvercetina (34,88 &mu;g/mL). U subkritičnom ekstraktu, dobijenom na optimalnim uslovima, detektovano je jedino prisustvo hidroksi metilfurfurala usled čega ovaj ekstrakt nije podvrgnut ispitivanju antimikrobne i antihiperglikemijske aktivnosti.<br />U cilju ekstrakcije lipofilnih komponenata M. vulgare primenjene su destilacija etarskog ulja (0,05%), Soxhlet ekstrakcija (3,23%) i ekstrakcija superkritičnim ugljendioksidom na pritiscima 100, 200 i 300 bar i temperaturama 40, 50 i 60 &deg;C. Najdominantnija komponenta u svim dobijenim superkritičnim ekstraktima je hemotaksonomski marker marubiin koji nije identifikovan u Soxhlet ekstraktu.<br />Tečni ekstrakti dobijeni ultrazvučnom i mikrotalasnom ekstrakcijom na optimizovanim uslovima i macerat su osu&scaron;eni spray drying tehnikom (uz dodatak 10 i 50% maltodekstrina) u cilju dobijanja stabilnije forme suvog ekstrakta. Suvim ekstraktima su određene najznačajnije fizičke osobine i hemijski sastav kao i antimikrobna i antihiperglikemijska aktivnost kako bi se procenila mogućnost njihove inkorporacije u funkcionalnu hranu, dijetetske suplemente ili u formulaciji različitih gotovih proizvoda koji se primenjuju preventivno ili radi pobolj&scaron;anja zdravlja ljudi.</p> / <p>Within the scope of this dissertation, the possibilities of applying modern methods for obtaining liquid (ultrasonic extraction, microwave extraction and subcritical water extraction) and lipophilic extracts (extraction with supercritical carbon dioxide) rich in polyphenols from the aboveground part of white horehound (Marrubium vulgare L.) were examined. Liquid extracts were optimized using an experimental design and response surface method, and subjected to spray drying. Liquid and dry extracts have been extensively examined for their chemical composition and biological activity, antimicrobial and in vitro antihyperglycaemic activities.<br />For each of these extraction techniques, the influence of the main parameters was examined: ultrasonic extraction (temperature, extraction time and ultrasound power), microwave extraction (ethanol concentration, extraction time and microwave power) and subcritical water extraction (temperature, extraction time and HCl concentration in the extractant). Liquid extracts obtained by ultrasonic and microwave extraction under optimized conditions revealed the presence of chlorogenic acid and quercetin, which were not detected in macerates. The antimicrobial activity of both extracts was examined and B. cereus was found to be the most sensitive bacterial strain, whereas in the case of yeast S. cerevisiae was particularly sensitive to both extracts. In addition to this activity, the in vitro antihyperglycaemic activities of both extracts against &alpha;-amylase and &alpha;-glucosidase were investigated, with ultrasound extract showing higher antihyperglycemic activity (50.63% inhibited &alpha;-amylase and 48.67% inhibited &alpha;-glucosidase) which could potentially be explained by the presence of chlorogenic acid (33.11 &mu;g/mL) and quercetin (34.88 &mu;g/mL). In the subcritical extract, obtained under optimal conditions, only the presence of hydroxy methylfurfural was detected, and this extract was not subjected to antimicrobial and antihyperglycemic activity.<br />For the extraction of lipophilic components of M. vulgare, distillation of essential oil (0.05%), Soxhlet extraction (3.23%) and extraction with supercritical carbon dioxide at pressures of 100, 200 and 300 bar and at temperatures of 40, 50 and 60 &deg; C were applied. The most dominant component in all the supercritical extracts obtained is the chemotaxonomic marker marubiin, which was not identified in the Soxhlet extract.<br />The liquid extracts obtained by the ultrasound and microwave extraction under optimized conditions and the macerate were spray dried (with the addition of 10 and 50% maltodextrin) to obtain a more stable form of the dry extract. Dry extracts underwent determination of the most important physical properties and chemical composition as well as antimicrobial and antihyperglycemic activities to evaluate their incorporation into functional foods, dietary supplements or in the formulation of various finished dosage forms that are administered preventively or to improve human health.</p>

<p><strong>Apstrakt je obrađen tehnologijama za optičko prepoznavanje teksta (OCR).</strong></p><p>Ispitana je ekstrakcija timijana (<em>Thymus vulgaris L.</em>) primenom ugljendioksida u superkritičnom stanju kao ekstragensa. Razrađen je postupak tečne hromatografije pod visokim pritiskom (HPLC) za određivanje farmakolo&scaron;ki aktivnih fenolnih jedinjenja, timola i karvakrola, koji obezbeđuje visok stepen reproduktivnosti određivanja (za timol K_w = 1,29%). Za kvalitativnu i kvantitativnu analizu proizvoda na bazi timijana, etarskog ulja i ekstrakata, primenjen je metod gasne hromatografije (GC) sa masenom spektrometrijom (MS). Sadržaj etarskog ulja određen oficinalnim postupkom koji propisuje jugoslovenska farmakopeja iznosi 1,75% (V/m). Dominantna komponenta etarskog ulja timijana je timol čiji je sadržaj 50,06% (m/m), dok je karvakrol prisutan, praktično, u tragovima (1,15%). Od prisutnih parafina u etarskom ulju, največi sadržaj ima n-tetradekan (16,11%). U totalnom ekstraktu timijana dobijenom metilen- hloridom primenom ekstrakcije sa povremenim ispu&scaron;tanjem ekstrakta (Ekstrakcija po Soxhlet-u), koji je dobijen u prinosu 4,95% (m/m), dominantna komponenta je n-tetradekan (47,18%), dok je sadržaj timola 16,11%.<br />Kinetika ekstrakcije timola se može uspe&scaron;no predstaviti analogno izrazu za kinetiku reakcije prvog reda, primenom koje se dobija največa vrednost konstante brzine ekstrakcije (k) 0,2209 h^-1 za selektovani protok superkritičnog ugijendioksida 97,725 dm³/h.<br />Primenom timijana stepena usitnjenosti d₂ srednjeg prečnika 0,35 mm za ekstrakciju superkritičnim ugijendioksidom (100 bar; 40&deg;C; 2,5 sata) postiže se kvantitativna ekstrakcije prisutnog etarskog ulja. Po svom sastavu ovaj ekstrakt je najsličniji etarskom ulju timijana dobijenom destliacijom pomoću vodene pare. Produžavanje vremena ekstrakcije iznad 2,5 sata ima za posledicu povećani prinos, pre svega, parafina.<br />Primenom ugljendioksida male moći rastvaranja (80 bar; 40⁰C;&nbsp; d=0,1918 g/cm³) ne postiže se kvantitativna ekstrakcija etarskog ulja. Povećavanjem radnog pritiska od 100 do 400 bar, usled povećanja moći rastvaranja, dobijaju se ekstrakti koji su po svom sastavu sve sličniji totalnom ekstraktu dobijenom ekstrakcijom metilenhloridom.<br />Za modelovanje ekstrakcionog sistema timijan - superkritični ugljendioksid je primenjena jednačina Reverchon-a i Sesti Osseo-a:<br />&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;Y = 100 [1-exp(-t/t_i)]<br />gde je: Y - normalizovani ekstrakcioni prinos (%); t - vreme ekstrakcije (s) i t_i -&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; vreme unutra&scaron;nje difuzije (s)<br />odnosno njen modifikovani oblik:<br />&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Y = 100 [1 - ehr (at + b)]<br />Izračunate vrednosti standardne gre&scaron;ke regresije (S_Y,X) ukazuju da u osnovi modifikovana jednačina bolje fituje normalizovane prinose kako totalnog ekstrakta, tako i timola, u odnosu na polaznu jednačinu.<br />Jednačine:<br />&nbsp;&nbsp; log S = a m_rel + b<br />i<br />&nbsp;&nbsp; log S = a <em>d</em>_C02 + b<br />gde je: S - rastvorljivost totalnog ekstrakta (g/dm³), odnosno timola (mg/dm ) u ugljendioksidu; m_rel - relativna masa ugljendioksida (g CO2/g timijana) i <em>d</em>_CO2 zapreminska masa ugljendioksida (g/cm³)<br />se mogu, takođe, uspe&scaron;no primeniti za modelovanje ekstrakcije ispitivanog sistema.<br />Primenom ugljendioksida u tečnom stanju (65 bar; 23&deg;C) se dobija ekstrakt timijana sličan po svom sastavu ekstraktu dobijenom ekstrakcijom superkritičnim ekstragensom na pritisku 100 bar.<br />U cilju dobijanja ekstrakta timijana koji je po svom kvalitativnom i kvantitativnom sastavu najsličniji etarskom ulju dobijenom oficinalnim postupkom destilacije pomoću vodene pare, neophodno je za ekstrakciju koristiti ugljendioksid zapreminske mase 0,6302 g/cm³( 100 bar;40&deg;C), biljni materijal stepena usitnjenosti d₂, vreme ekstrakcije 2,5 sata i protok ekstragensa 97,725 dm³/h.</p> / <p><strong>Abstract was processed by technology for Optical character recognition (OCR).</strong></p><p>The extraction of thyme (Thymus vulgaris L.) by supercritical carbon dioxide as an extragent was investigated. The developed HPLC (High Pressure Liquid Chromatography) method for determination of pharmacologically active phenolic compounds, thymol and carvacrol, showed a high reproducibility (Kw for thymol 1.29%). Qualitative and quantitative analyses of thyme products, essential oil and extracts, were carried out by gas chromatography - mass spectrometry (GC-MS). The content of essential oil determined by an official procedure given by Yugoslav pharmacopoeia was 1.75% (V/w). The predominant compound of the essential oil is thymol (50.06%; w/w), while carvacrol is present, practically, in traces (1.15%; w/w). The content of n-tetradecane in the thyme essential oil (16.11%; w/w) is higher than that of other paraffins. In the total thyme extract obtained by methylene chloride using extraction with periodical exchange of the extract (Soxhlet extraction) in a yield of 4.95%(w/w), the predominant compound was n-tetradecane (47.18%), while the content of thymol was 16.11%.</p><p>The extraction kinetic of thymol can be successfully expressed by analogy to first order reaction kinetic, and a highest value of extraction rate constant (k) of 0.2209 h-1 for the selected flow rate of supercritical carbon dioxide of 97.725 dm3/h.<br />A quantitative extraction of thyme essential oil by supercritical carbon dioxide (100 bar; 40&deg;C; 2.5hours) was obtained using thyme of the grinding degree d2 (mean particle diameter 0.35 mm). The composition of this extract is very similar to that of the essential oil obtained by steam distillation. A higher yield of extraction, primarily of paraffins, was obtained by prolonging the extraction time over 2.5 hours. The use of carbon dioxide of low solubility power (80 bar; 400C; d =0.1918 g/cm3) did not yield a quantitative extraction of thyme essential oil.<br />Ву increasing extraction pressure from 100 to 400 bar, i.e. solubility power, the extract composition was more and more similar to the composition of total extract obtained by methylene chloride.<br />То model extraction of the system thyme - supercritical carbon dioxide use was made of the Reverchon - Sesti Osseo equation:</p><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Y = 100 [1-exp(-t/ti)]</p><p>where: Y - normalized extraction yield (%); t - extraction time (s) and ti - internal diffusion time (s)<br />i.e. of its modified form:<br />&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Y = 100 [1 - ехр (at + b)]<br />The calculated values of standard error of regression (SY,X) show that the modified equation is generally a better fit for the normalized yields of total extract and thymol compared to the original equation.<br />Equations:</p><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; log S = a mrel + b<br />and<br />&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; log S = a dC02 + b<br />where: S - solubility of total extract (g/dm3), i.e. of thymol (mg/dm3) in carbon dioxide; mrel - relative mass of carbon dioxide (g CO2/g thyme) and dCO2 carbon dioxide density (g/cm3)</p><p>could be successfully used for modelling of the investigated extraction system.<br />The composition of thyme extract obtained by liquid carbon dioxide (65 bar; 23&deg;C ) is very similar to that obtained by supercritical extragent at a pressure of 100 bar.<br />То obtain a thyme extract whose qualitative and quantitative characteristics are most similar to those of the essential oil obtained using official procedure by steam distillation, it is necessary to use a carbon dioxide density of 0.6302 g/cm3 (100 bar; 400C), grinding degree of thyme d2, extraction time of 2.5 hours and extragent flow rate of 97.725 dm3/h.</p>

<p style="text-align: justify;">U okviru ove doktorske disertacije ispitana je mogućnost ekstrakcije bioaktivnih jedinjenja iz biljne pra&scaron;ine A. uva-ursi, koja nastaje kao sporedni proizvod u fabrici filter čaja. Za ekstrakciju bioaktivnih jedinjenja primenjene su različite tehnike ekstrakcije (klasična ekstrakcija, ultrazvučna ekstrakcija i ekstrakcija subkritičnom vodom), kao i različiti ekstragensi: voda na sobnoj temperaturi, 30-70% etanol i subkritična voda. Sadržaj bioaktivnih jedinjenja u ekstraktima ispitan je primenom odgovarajućih spektrofotometrijskih metoda i HPLC metode. U ekstrakciji biljne pra&scaron;ine A. uva-ursi detaljno je matematički analiziran uticaj procesnih parametara na ekstrakciju subkritičnom vodom bioaktivnih jedinjenja iz materijala ovog tipa i u tom smislu je posmatran uticaj temperature, vremena i dodatka kiseline, a takođe su određeni uslovi pri kojima je, primenom ove tehnike, moguće ostvariti maksimalnu ekstrakciju bioaktivnih jedinjenja od značaja. Pored toga prikazana je i mogućnost primene spray drying tehnike za su&scaron;enje tečnih i proizvodnju suvih ekstrakta A. uva-ursi u formi praha. Na kraju, na osnovu koncentracije bioaktivnih jedinjenja i njihovog delovanja, dat je predlog nekoliko novih preparata na bazi ekstrakata A. uva-ursi, u tečnoj i čvrstoj formi.</p> / <p>This dissertation investigated the possibility to extract bioactive compounds from A. uva-ursi herbal dust, which occurs as a by-product at the filter-tea<br />factory. Various extraction techniques (classical extraction, ultrasound assisted extraction and subcritical water extraction), were applied for the extraction of bioactive compounds, as well as different solvents: water at room temperature, 30-70% ethanol and subcritical water. The content of bioactive compounds in the extracts was investigated using appropriate spectrophotometric methods and the HPLC method. In the extraction of A. uva-ursi herbal dust a detailed mathematical analysis of the influence of process parameters on subcritical water extraction of bioactive compounds from this type of material was conducted. In this regard the influence of temperature, time and acid addition was observed and the conditions under which it is possible to achieve maximum extraction of significant bioactive compounds, by applying this technique, were determined. In addition, the possibility of applying the spray drying technique for drying the liquid and producing dry A. uva-ursi extracts in the form of powder is also presented. Eventually, based on the concentration of the bioactive compounds and their activity, a proposal for a several new preparations in liquid and solid form, based on A. uva-ursi extracts is given.</p>

<p>&nbsp;Upotrebom različitih ekstragenasa izvr&scaron;ena je ekstrakcija ploda, lista i korena belog (<em>Morus alba</em> L.) i crnog duda (<em>Morus nigra</em> L. ). Ekstrakcija etanolom je izvr&scaron;ena pri predhodno određenim optimalnim uslovima ekstrakcije (70% koncentracija etanola, temperatura 60&deg;C i odnos rastvarač:droga 20 ml/g). Ispitan je hemijski sastav ekstrakata različitih delova duda. Ispitivanja hemijskog sastava etanolnih ekstrakata obuhvatala su određivanje sadržaja ukupnih fenola, ukupnih flavonoida, ukupnih i monomernih antocijana, kvalitativni i kvantitativni sastav polifenolnih komponenata, kao i sadržaj mikro-, makro- i toksičnih elemenata. Najvedi sadržaj ukupnih fenolnih jedinjenja određen je u ekstraktu korena <em>M. nigra</em> (186 mg EHK/g), a najvedi sadržaj flavonoidnih jedinjenja u ekstraktu lista <em>M. nigra</em> (67 mg ER/g). U ekstraktima lista duda po svom sadržaju u odnosu na ukupan sadržaj polifenolnih jedinjenja ističe se kafena kiselina (od 53,34 do 62,11%), dok se u ekstraktima korena izdvaja hlorogenska kiselina (udeo vedi od 74%). U pogledu mineralnog sastava ekstrakata relativno veliki sadržaj Ca i Mg je karakterističan za obe vrste ekstrakata (vedi od 1.400 &mu;g/g). Mikroelementi u svim ispitanim uzorcima duda po opadajudem nivou, su Fe, B, Zn, Cu i Mn. U ekstraktima dobijenim superkritičnim CO₂određen je sadržaj masnih kiselina i ukupnih karotenoida. U ispitivanim ekstraktima dominante su nezasidene masne kiseline, linolna i linolenska kiselina, a značajan je udeo i zasidene masne kiseline (palmitinske kiseline).</p><p>Ispitivanje delovanja ekstrakata obuhvatalo je evaluaciju antioksidativne aktivnosti pradenjem sposobnosti neutralizacije slobodnih radikala, redukcionog potencijala i inhibicije lipidne peroksidacije. Po svim primenjenim metodama najvedu aktivnost je pokazao ekstrakt lista crnog duda. U zavr&scaron;noj fazi rada ispitana je in vitro citotoksična aktivnost ekstrakata duda, njihovim delovanjem na rast tri delijske linije: Hep2, RD i L2OB. Ekstrakti lista <em>M. nigra</em> su se pokazali kao najpotentniji inhibitori&nbsp;rasta na sve tri delijske linije (IC₅₀&lt;30 &mu;g/g). Na osnovu svih sprovedenih ispitivanja kao proizvod sa najboljim karakteristikama pokazao se ekstrakti lista <em>M. nigra</em>.</p> / <p>Using different extraction solvents extracts of fruit, leaves and roots of white (<em>Morus alba</em> L.) and black mulberry (<em>Morus nigra</em> L.) were prepared. Extraction by ethanol was carried out at the previously determined optimal extraction conditions (70% ethanol concentration, temperature 60&deg;C and the ratio of solvent: drug 20 ml/g). The chemical composition of extracts obtained from different parts of the mulberry were analysed. These analyses included the determination of total phenols, total flavonoids, monomeric and total anthocyanins, qualitative and quantitative composition of polyphenolic components, and the content of micro-, macro- and toxic elements. The highest phenolic content was determined in the extract of <em>M. nigra</em> root (186 mg ECA/ g), while the highest content of flavonoid compounds was determined in the extract of <em>M. nigra</em> leaves (67 mg ER/g). In the case of mulberry leaf extracts dominant phenolic compound, in relation to the total phenolic content, was caffeic acid (with contribution from 53.34 to 62.11%), while in the extracts of root extracts dominant phenolic compounds was chlorogenic acid (a higher proportion of 74%). In terms of mineral composition of the extracts relatively high content of Ca and Mg was characteristic of both types of extracts (greater than 1,400 mg/g). Measured content of trace elements in mulberry extracts decrease in the next order: Fe&gt; B&gt; Zn&gt; Cu&gt; Mn. In the extracts obtained by supercritical CO₂ fatty acids and total carotenoids were determined. In these extracts dominant unsaturated fatty acids were linoleic and linolenic acid. Also, significant proportion of saturated fatty acid (palmitic acid) has been detected.</p><p>Testing of extracts activity included the evaluation of antioxidant activity applying the methods based on: neutralize free radicals, reducing power potential and the inhibition of lipid peroxidation. All applied methods showed the highest activity of black mulberry leaf extract. In the final phase of work in vitro cytotoxic activity of mulberry extracts was examined. Their effect on growth of three cell lines: Hep2, RD, and L2OB were analysed. Extracts of <em>M. nigra</em> proved to be the most potent inhibitors of growth in all three cell lines (IC₅₀&lt;30 mg/g). On the basis of all conducted tests it has been proven that <em>M. nigra</em> extracts are products with the best characteristics.</p>

Augant pasaulinei pramonei aplinkos tarša nepaliaujamai didėja, tad augalai augantys užterštoje aplinkoje, savo sudėtyje taip pat turi nemažai įvairių junginių, kurie gali įtakoti juos vartojančių žmonių sveikatą. Eterinių aliejų išgautų iš augalų tyrimas yra vienas iš būdų nustatant augalo kokybę, užtikrinant, kad taršalai nepateks tolesniam perdirbimui. Eteriniams aliejams išgauti yra daug būdų, kaip kad distiliacija vandens garais, šviežių aromatinių dalių išspaudimas, ekstrakcija tirpikliais, tačiau tokie išgavos būdai būdingesni išgaunant didelius kiekius, tad reikia ir daug žaliavos. Tyrimams naudojami greitesni, bei mažiau žaliavos reikalaujantys metodai, kaip, kad ekstrakcija superkritiniais skysčiais, kietafazė mikroekstrakcija bei ekstrakcija iš bandinio viršerdvės. Šio darbo tikslas - eterinių aliejų, išgautų superkritinių skysčių ekstrakcijos, kietafazės mikroekstrakcijos bei tiesioginės viršerdvės ekstrakcijos būdais, sudėties nustatymas dujų chromatografijos – masių spektroskopijos metodu. Panaudojant gautus duomenis nustatyti kuris ekstrakcijos būdas pasižymi didžiausiu efektyvumu. Duomenų tikslumo užtikrinimui bandiniai pakartotinai leisti po 3 kartus, identifikuoti junginiai tikrinti skaičiuojant kovačo retencijos indeksus ir juos lyginant su jau žinomais indeksais internetinėje bibliotekoje. Darbo metu remiantis gautais duomenimis nustatyta efektyviausias eterinių aliejų ekstrakcijos metodas, junginiai, esantys bandiniuose bei būdingųjų junginių kiekis. / As the word industry develops, the amount of wastes, that are being released in to our environment is always increasing, as a result vegetables that are growing in such a environment are also contaminated with compounds that can affect the health of people that are using them. Many methods can be used to obtain essential oils, such as distillation using water vapour, extruding from fresh fragrant, extraction using solvents, but these kind of methods are used for getting large amounts of essential oils, so staple is used. For chemical analysis faster and less staple demanding methods are used, such as, super critical fluid extraction, solid phase micro extraction or direct head space extraction. The goal of this work is to determine the composition of essential oils and to estimate witch, super critical fluid extraction, solid phase micro extraction or direct head space extraction is the best for obtaining essential oils from spice herbs. In order to secure the precision of data each sample was analyzed three times, for correct compound identification kovach retention indexes were calculated and compared with the known ones on the internet library. Using the obtained data the most effective method foe essential oils obtaining was determined, also compound composition in samples and the amount of specific compounds were analyzed.

<p>Dokazivanje virusnih infekcija pĉela od velikog je interesa za pravilnu procjenu odnosa izmeĊu domaćina i potencijalnog patogena. Do izrade ove doktorske teze, na teritoriji Bosne i Hercegovine nije bilo prouĉavanja koja bi dokaz virusnih infekcija pĉela dovela do mogućnosti kliniĉke procjene patolo&scaron;kog odnosa za eko-genotip prilagoĊene pĉelinje zajednice. Evolucijski adaptirani virusi medonosne pĉele Apis mellifera carnica, sa stanovi&scaron;ta veterinarske entomologije, veterinarske virusologije i infektivnih bolesti, te saznanja veterinarskog servisa uop&scaron;te, nisu bili dovoljno prepoznati. Pojava i dostupnost molekularnih dijagnostiĉkih metoda omogućile su senzitivno i specifiĉno dokazivanje dijela genoma virusa koji inficiraju pĉelinje zajednice. Iz ekstrahovanih uzoraka templata razliĉitih uzoraka homogenizata sluĉajno uzorkovanih pĉela, RNK, sa jednim parom prajmera za svaki virus, pojedinaĉno, redoslijedom: BQCV, DWV, SBV, ABPV, KBV, CBPV uraĊeni su amplifikacijski protokoli i dobijeni rezultati za klasiĉan, end point, RT-PCR. Ovom doktorskom disertacijom dobijeni su prvi rezultati koji se odnose na utvrĊivanje prisustva i ra&scaron;irenosti virusnih infekcija kod vrste Apis mellifera u pĉelinjacima Bosne i Hercegovine. Od oko dvadeset virusa koji inficiraju pĉele i pĉelinje zajednice, metodama primjenjive molekularne dijagnostike u ovoj doktorskoj tezi dokazivali smo prisustvo virusnih infekcija za &scaron;est najvi&scaron;e prouĉavanih virusa pĉela. Dobijeni rezultati dokazali su prisustvo pet od &scaron;est traţenih virusa sa razliĉitim kombinacijama koinfekcija te raznolikom prostornom distribucijom virusa. Poznavanje sloţenih mehanizama virusnih infekcija pĉelinjih zajednica neophodnost je koja sa novim saznanjima progla&scaron;ava neke od virusa emergentnim patogenima, koji bitno naru&scaron;avaju opstanak pĉelinje zajednice. U patolo&scaron;ko-sinergistiĉkom odnosu sa nametnikom pĉelinje zajednice Varroa destructor neki pĉelinji virusi uzroci su globalno registrovanih gubitaka. Dio istraţivanja u radu odnosio se na terensko prouĉavanje dinamike prisustva grinje Varroa destructor u pĉelinjim zajednicama. Za pravilno dono&scaron;enje suda o uzroku ugibanja pĉelinjih zajednica, pored postojećih zahtjeva kontrole patogena pĉela definisanih procedurama dijagnostiĉkog manuala i Koda OIE-a, bilo bi potrebno imati i rezultate kojim bi se dokazalo kvalitativno i kvantitativno prisustvo virusa kod vrste Apis mellifera. UvoĊenje i standardizacija molekularno-biolo&scaron;kih metoda u rutinskoj dijagnostici virusnih bolesti pĉela je opravdano. Samo je pitanje vremena kada će virusne infekcije pĉela biti predmet rutinske dijagnostike kojim se kontroli&scaron;ezdravstveni status pĉelinje zajednice.</p> / <p>Evidence of viral infection of bees is of great interest for the proper assessment of the relationship between host and pathogen potential. By making this doctoral thesis on the territory of Bosnia and Herzegovina, where before was no present study of evidence of viral infections, lead to ability on clinical evaluation of pathological relations for eco-genotype adapted colonies. Evolutionarily adapted viruses honeybee Apis mellifera, from the veterinary entomology, veterinary virology and infectious diseases, and knowledge of the veterinary services in general, have not been recognized enough. The appearance and availability of molecular diagnostic methods have allowed sensitive and specific detection part of the genome of the virus that infects the colonies. Templates were extracted from samples of different patterns, homogenisate, randomly sampled bees, RNA, with a pair of primers for each virus, respectively, in the order: BQCV, DWV, SBV, ABPV, KBV, CBPV amplification protocols were done and the results obtained by conventional, end point, RT -PCR. This doctoral dissertation obtained the first results concerning the determination of the presence and spread of viral infections in the species Apis mellifera in Bosnia and Herzegovina. From about twenty viruses that can infect bees and hives using diagnostic methods applicable in this doctoral thesis we demonstrate the presence of viral infections of six most studied viruses of bees. The obtained results showed the presence five of the six essential viruses in forms of co-infection with different combinations and varied virus spatial distribution. Awareness of the complex mechanisms of viral infections in colonies is that the new knowledge proclaimed a virus as &quot;emergent pathogen&quot; that significantly impairs the survival of honeybee colonies. The pathogenic synergistic relationship with the parasite Varroa destructor causes that bee viruses are globally listed as reasons for losses. Part of the research work was related to the field study of the dynamics of the presence of Varroa destructor mites in honey bee colonies. For a accurate judgment of the deaths of bee colonies, in addition to existing requirements bee pathogen control procedures defined by diagnostic Manuals and Code of the OIE, it would be necessary to have the results to prove the qualitative and quantitative presence of the virus in the species Apis mellifera. The introduction and standardization of molecular biological methods in routine diagnosis of viral diseases of bees is justified. It is only a matter of time before the virus infections of bees will be subject to routine diagnostics which controls the health of bee colonies</p>

Evaluation of solid-phase microextraction as an alternative official method for analysis of polymers migration. The objective this study was to compaire the official methods with solid-phase microextraction (SPME)for the analysis of compounds migrating from cross-linked polyethylene into water. A medium polarity polydimethylsiloxane/divinylbenzene (PDMS/DVB)was proved most efficient for the SPME extraction. However, when applied to water samples in contact with polyethylene, SPME proved to be immensely more sensitive and have a greater extraction range than liquid-liquid extraction (LLE). It was proved the migration of Phenol, 2,4-bis(1,1-dimethylethyl). Concentration of this compound 0,6-0,15mg/l.It was proved the migration of Cyclohexadiene-1,4-dione, 2,6-bis(1,1-dimethylethyl).

Tyrimo objektas – kraujo plazma su vaistinių medžiagų mišiniu (buspirono hidrochloridu ir fluoksetino hidrochloridu). Šio tyrimo tikslas – nustatyti tinkamiausias ekstrakcijos sąlygas, reikalingas vaistų mišinio (buspirono ir fluoksetino) išskyrimui iš kraujo plazmos bei pritaikyti efektyviosios skysčių chromatografijos metodiką vaistų mišinio veikliųjų junginių identifikavimui ir kiekio nustatymui. Darbo uždaviniai buvo pritaikyti ir validuoti efektyviosios skysčių chromatografijos metodiką; išskirti buspirono ir fluoksetino vaistų mišinį iš kraujo plazmos skysčių – skysčių (SSE) ir kietafazės ekstrakcijos (KFE) metodais; optimizuoti geriausią ekstrakcijos metodą. Tyrimo metu buvo pritaikyta ESC metodika buspirono ir fluoksetino identifikavimui ir kiekio nustatymui. Atlikta ESC metodikos validacija: įrodytas specifiškumas, rezultatų glaudumas, remiantis koreliacijos koeficientu (R2) įvertintas metodikos teisiškumas, buspirono ir fluoksetino koreliacijos koeficientai atitinkamai lygūs 0,9997 ir 0,9998. Nustatytos aptikimo ribos (buspironui 0,4 µg/ml, fluoksetinui 0,75 µg/ml) ir nustatymo ribos (buspironui 0,75 µg/ml, fluoksetinui 1 µg/ml). Vaistinių medžiagų mišinys išskirtas iš kraujo plazmos SSE ir KFE metodais. Nustatyta, kad KFE yra greitesnis ir tikslesnis metodas lyginant su SSE, todėl KFE pasirinkta tolesniam metodo optimizavimui. Eksperimentai atlikti su 6 organiniais tirpikliais (metanoliu, etanoliu, propanoliu, trichlormetanu, dichlormetanu ir acetonitrilu)... [toliau žr. visą tekstą] / The object of study – human plasma with the mix of two drugs (buspirone hydrochloride and fluoxetine hydrochloride). The aim of this study was to determine extraction conditions for buspirone and fluoxetine isolation from human plasma and to apply high-performance liquid chromatography (HPLC) method for quantitative analysis of the drugs after extraction. The objective was to apply and validate HPLC procedure; to extract buspirone and fluoxetine from human plasma by liquid-liquid extraction (LLE) and solid phase extraction (SPE); to optimize efectiveness extraction method. The HPLC method for identification and quantitative analysis of drugs was optimized. The mobile phase was 0,1 per cent of trifluoroacetic acid and acetonitrile.Validation of the HPLC method was carried out. The specificity, precision, linearity, limits of detection (buspirone 0,4 µg/ml, fluoxetine 0,75 µg/ml) and quantification (buspirone 0,75 µg/ml, fluoxetine 1 µg/ml) were determined. Validate HPLC method was applied for analysis of buspirone and fluoxetine after extraction. Drugs were extracted from human plasma by LLE and SPE methods. The best recovery of analites gave SPE methods. The recoveries of the drugs using six different organic solvents (methanol, propanol, ethanol, trichloromethane, dichloromethane, acetonitrile) were examined. Selected the most appropriate environment (acidify) for methanol and the methanol percentage of the elution solvent (80 per cent). Selected two of the most appropriate... [to full text]

<p>Predmet izučavanja ove disertacije je ispitivanje mogućnosti imobilizacije&nbsp;toksičnih metala u sedimentu primenom imobilizacione tehnike&nbsp;solidifikacije/stabilizacije portland cementom, kalcijum-oksidom, prirodnim zeolitom,&nbsp;letećim pepelom, kaolinitom, montmorilonitom, i različitim sme&scaron;ama ovih agenasa&nbsp;kao i određivanje njihove efikasnosti u zavisnosti od brojnih faktora. Efikasnost&nbsp;imobilizacionih postupaka praćena je kori&scaron;ćenjem tzv. testova izluživanja, odnosno&nbsp;određivanjem koncentracije metala koji su u toku određenog vremena iz imobilisane&nbsp;faze pre&scaron;li u mobilnu fazu gde se smatraju potencijalno opasnim i biodostupnim.&nbsp;Krajnji cilj svakog testa izluživanja je mogućnost procene adekvatnosti primene&nbsp;određene imobilizacione, odnosno remedijacione tehnike. Efikasnost imobilizacionih&nbsp;postupaka zavisi i od određenih osobina: pH rastvora, dodatka imobilizacionog agensa&nbsp;&scaron;to je istraživanjem ispitano. Na osnovu ovog testa, odnosno &nbsp; odnosa kumulativne&nbsp;frakcije metala koja je oslobođena iz imobilizacione sme&scaron;e u mobilnu fazu i vremena&nbsp;određen je tip mehanizma koji omogućava transport metala &nbsp;(spiranje, difuzija,&nbsp;rastvaranje). Takođe su određeni i i parametri (koeficijenti difuzije, indeksi&nbsp;izlužljivosti) koji&nbsp; će poslužiti za ocenu efikasnosti prethodno primenjenih&nbsp;imobilizacionih tehnika. Primenjeni su i modifikovani testovi izluživanja sa ciljem &scaron;to&nbsp;bolje simulacije realnih uslova. U modifikovanim testovima izluživanja kori&scaron;ćen je&nbsp;rastvor sa pH 3,25 da bi se simulirali uslovi koji opona&scaron;aju kisele ki&scaron;e, odnosno uslovi&nbsp;u realnom sistemu. Takođe je kor&scaron;ćen i rastvor huminskih materija sa ciljem&nbsp;simulacije uslova velikog organskog opterećenja do kojeg bi moglo doći u slučaju&nbsp;akcidentnih situacija (na primer poplava) i generalno uslova koji se stvaraju u &nbsp;prirodi&nbsp;pri raspadanju organskog materijala (li&scaron;će, trava, itd.). Rezultati dobijeni simulacijom&nbsp;ovih uslova će dalje omogućiti modelovanje pona&scaron;anja metala u smislu dugoročnog&nbsp;&quot;izluživanja&quot; iz tretiranog sedimenta kao i procenu najefikasnijih agenasa &nbsp; za&nbsp;imobilizaciju različitih metala u sedimentu. Na osnovu dobijenih rezultata, zaključeno&nbsp;je da su optimalni agensi za imobilizaciju metala u sedimentu sme&scaron;a cementa i kreča&nbsp;(5% cementa i 10% kreča), sme&scaron;a montmorilonita i kreča (30% montmorilonita i 10%&nbsp;kreča), leteći pepeo (30%) i zeolit (30%). U ovim sme&scaron;ama dominantan mehanizam&nbsp;izluživanja je difuzija, a ove sme&scaron;e se mogu smatrati inertnim otpadom po svim&nbsp;ispitivanim kriterijumima. Ovi rezultati se mogu upotrebiti za projektovanje i&nbsp;izgradnju pilot postrojenja na kome bi se ispitala efikasnost ovih agenasa za&nbsp;remedijaciju sedimenta u realnim uslovima. Dobijeni podaci su neprocenjivi sa&nbsp;aspekta ekonomski i ekolo&scaron;ki prihvatljivog upravljanja sedimentom.</p> / <p>This work is concerned with exploring the possibilities of immobilization of toxic&nbsp;metals in sediments using solidification/stabilization as imobilization technique and&nbsp;using Portland cement, calcium oxide, natural zeolite, flying ash, kaolinite, montmorilonite, and various mixtures of these agents and determine their&nbsp;effectiveness depending on many factors. Performance of imobilization procedures&nbsp;was followed by the use of so-called leaching tests and determination of the metals&nbsp;concentration that are within a specified interval of time leached from the immobile phase and as such can be considered potentially hazardous and bioavailable. The ultimate goal of every leaching test is to assess the adequacy of the possibility of applying certain imobilization or remediation technique. Performance of imobilizaction procedures depends on certain characteristics: pH of solution, additon of imobilization agent, etc. Based on this test, ie relation between the cumulative fraction of metal leached from imobilization mixture and time, the type of leaching mechanism that allows the transport of metals (wash-off, diffusion, dissolution) was determined. Other parameters which will serve for evaluating the efficiency of the previously applied imobilization techniques (diffusion coefficients, leaching indices) were also determined. Modified leaching tests were applied with the aim of better simulation of real conditions on the field. In the modified leaching tests the solution with pH 3.25 was used to simulate conditions that mimic acid rain and conditions in the real system. The humic acid solution was also used with the aim to simulate high organic loads which could occur in the case accident situation (eg floods) and the general conditions that are created in naturewhen decaying organic material (leaves, grass, etc.). The results obtained by simulating these conditions will enable the modeling of behavior of metals in terms of long-term leaching period from the treated sediment and assess the most effective agents for the immobilization of various metals in the sediments. Based on these results, it was concluded that the optimal agents for immobilization of metals in the sediment mixture of cement and lime (5% cement and 10% lime), a mixture montmorilonite and&nbsp; lime (30% montmorilonite and 10% lime), fly ash (30%) and zeolite (30%). In these mixtures dominant leaching mechanism is diffusion, and these mixtures can be considered as inert waste by all tested criteria. These results can be used to design and builda pilot plant in which order to evaluate the effectiveness of these agents for remediation of sediment in real &nbsp;terms. The obtained data are invaluable from the aspect of economic and ecologically acceptable management of sediment.</p>

<p>U okviru ove doktorske disertacije ispitane su i<br />poređene različite ekstrakcione tehnike korena<br />gaveza, lista crnog duda i peteljki tre&scaron;nje. Poređene<br />su konvencionalne i savremene ekstrakcione<br />tehnike i izvedena je optimizacija ekstrakcionih<br />postupaka s ciljem dobijanja ekstrakata ispitivanih<br />biljnih vrsta sa najvećom bioaktivno&scaron;ću. Za svaku<br />ekstrakcionu tehniku ispitan je uticaj najznačajnijih<br />parametara: maceracija (rastvarač i vreme<br />ekstrakcije), ubrzana ekstrakcija (temperatura i<br />rastvarač), supekritična ekstrakcija (pritisak i<br />koncentracija ko-rastvarača) i ekstrakcija<br />subkritičnom vodom (temperatura, pritisak, vreme<br />ekstrakcije i brzina me&scaron;anja). Ekstrakcija polifenola<br />korena gaveza, lista crnog duda i peteljki tre&scaron;nje je<br />bila najefikasnija ekstrakcijom subkritičnom<br />vodom. Primenjene su tehnika visokopritisne tečne<br />hromatografije spregnute sa masenom<br />spektrometrijom uz primenu različitih masenih<br />analizatora, masenog analizatora na bazi vremena<br />preleta jona i tandema kvadrupolnog i analizatora<br />na bazi vremena preleta jona. Za karakterizaciju<br />dobijenih ekstrakata instrumentalnom analizom su<br />određene kvalitativne i kvantitativne karakteristike dobijenih ekstrakata korena gaveza, lista crnog duda i peteljki tre&scaron;nje. HPLC-ESI-QTOF-MS/MS analizom ekstrakata identifikovano je vi&scaron;e od 27 novih jedinjenja različitih hemijskih klasa u svakom od ispitivanih biljnih matriksa.</p> / <p>Within the scope of this doctoral dissertation,<br />different extraction techniques of comfrey root,<br />black mulberry leaves and sweet cherry stems were<br />applied and compared. Conventional and novel<br />extraction techniques were optimized and<br />compared in order to obtain plant extracts with the<br />highest content of bioactive compounds. The<br />influence of the most important parameters was<br />investigated for each extraction technique:<br />maceration (solvent and extraction time),<br />accelerated solvent extraction (temperature and<br />solvent), supercritical fluid extraction (co-solvent<br />percentage and pressure) and subcritical water<br />extraction (temperature, pressure, extraction time<br />and agitation rate). Extraction of polyphenolic<br />compounds from comfrey root, black mulberry<br />leaves and sweet cherry stems was the most<br />effective using subcritical water. High-pressure<br />liquid chromatography coupled to mass<br />spectrometry techniques using time-of-flight and<br />quadrupole-time-of-flight mass analyzers have<br />been used to characterize bioactive compounds.<br />Qualitative and quantitative characteristics of the<br />obtained comfrey root, black mulberry leaves and sweet cherry stem extracts were determined. HPLC-ESI-QTOF-MS/MS revealed the presence of more than 27 compounds of different chemical classes that have been identified for the first time in&nbsp; the plant matrix.</p>